تعیین تراکم جمعیت Aspergillus flavus و Aspergillus parasiticus در تیمارهای آخرین آبیاری قبل از برداشت در خاک و میوه درختان پسته

نوع مقاله : مقاله پژوهشی

نویسندگان

1 پژوهشکده پسته، موسسه تحقیقات علوم باغبانی، سازمان تحقیقات، آموزش و ترویج کشاورزی، رفسنجان، ایران

2 بخش تحقیقات گیاهپزشکی، مرکز تحقیقات کشاورزی، منابع طبیعی و آموزشی استان سمنان (شاهرود) سازمان تحقیقات، آموزش و ترویج کشاورزی،

چکیده

هدف: آلودگی پسته به قارچ Aspergillus و آفلاتوکسین یکی از مهمترین مشکلات در تولید و صادرات این محصول با ارزش، می­باشد. این  پژوهش با هدف شناسایی دو گونه Aspergillus flavus و A. parasiticus، تعیین فراوانی جدایه­های توکسین­زا و غیرتوکسین­زای این دو گونه و جمعیت Aspergillus flavus clade در خاک، میوه­های سالم و ترک­خورده رقم اوحدی، در شش تیمار آخرین آبیاری قبل از برداشت (5، 10، 15، 20، 25 و 30 روز) اجرا گردید.
مواد و روش‌ها: جداسازی قارچ­های Aspergillus flavus clade از خاک، میوه­های سالم و ترک­خورده با روش تهیه سوسپانسیون و سری­های رقت، روی محیط کشت DRBC انجام شد. شناسایی دو گونه Aspergillus flavus و
A. parasiticus با استفاده از ویژگی­های ماکرومورفولوژیکی و مولکولی (پرایمر کالمودولین) و تشخیص و غربالگری جدایه­های توکسین­زا با استفاده از دو روش محیط کشت نارگیل-آگار (Coconut agar medium=CAM) و کروماتوگرافی لایه نازک (TLC) انجام شد. آزمایش در قالب طرح بلوک کامل تصادفی با چهار تکرار در طی سال­های 1392 تا 1394 اجرا گردید.
نتایج: از مجموع 233 جدایه Aspergillus flavus clade جمع­آوری شده، به ترتیب 221 و 12 جدایه متعلق به دو گونه Aspergillus flavus و A. parasiticus بودند. در میوه­های ترک­خورده، سالم و خاک فراوانی جدایه­های توکسین­زای
A. flavus به ترتیب 5/86، 5/87، 7/88 درصد و جدایه‏های توکسین‏زای A. parasiticus،  80، 75، 100 درصد بود. بر اساس نتایج تجریه مرکب، تیمارهای آخرین آبیاری 30 و 25 روز قبل از برداشت (تیمارهای با کمترین رطوبت خاک) کمترین و تیمارهای آخرین آبیاری 10 و 5 روز قبل از برداشت (تیمارهای با بیشترین رطوبت خاک) بیشترین جمعیت Aspergillus flavus clade را در خاک، میوه­های سالم و ترک خورده نشان دادند. استفاده از کود دامی جمعیت Aspergillus flavus clade را به طور معنی­داری افزایش داد به طوری که این افزایش در میوه­های ترک خورده، سالم و خاک در سال 1393 به ترتیب 66، 83 و 114 درصد بود.
نتیجه‌گیری: نتایج تحقیق حاضر نشان داد که کاهش فاصله آخرین آبیاری با زمان برداشت میوه (بالا بودن درصد رطوبت خاک سطحی باغ) و استفاده از کود دامی می­تواند موجب افزایش جمعیت Aspergillus flavus clade در خاک، میوه­های سالم و ترک­خورده پسته گردد. با توجه به اینکه افزایش جمعیت قارچی همراه با فراوانی جدایه­های توکسین­زای دو گونه Aspergillus flavus و A. parasiticus می باشد با تنظیم دور آبیاری، به طوری که آخرین آبیاری 25 تا 30 روز قبل از برداشت میوه باشد، می­توان آلودگی میوه­های پسته به Aspergillus و تولید آفلاتوکسین را کاهش داد.

کلیدواژه‌ها

عنوان مقاله [English]

Determination of population density of Aspergillus flavus and Aspergillus parasiticus in the last irrigation treatments before harvest in soil and fruits of pistachio trees

نویسندگان [English]

  • Amir hossein Mohammadi 1
  • Naser Sedaghati 1
  • Masoumeh Haghdel 1
  • Seyd Javad Hosseinifard 1
  • Mehdi Mohammadi Moghadam 2
1 Assistant Professor, Pistachio Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Rafsanjan, Iran.
2 Assistant Professor, Horticultural and Crop Sciences Research Department, Agricultural and Natural Resources Research and Education Center of Semnan Province (Shahrood), AREEO, Shahrood, Iran.
چکیده [English]

Objective
Contamination of pistachio with Aspergillus and aflatoxin is one of the most important problems in the production and export of this valuable product. In this research, in addition to molecular identification of Aspergillus flavus and A. parasiticus and determination of the frequency of their toxigenic and atoxigenic isolates, the population of Aspergillus flavus clade was measured in soil, intact and cracked fruits of cv. Ohadi in 6 treatments of last irrigation before harvest (5, 10, 15, 20, 25 and 30 days).
 
Materials and methods
Isolation of Aspergillus flavus clade fungi from soil, intact and cracked fruits was done by serial dilution method on DRBC culture medium. Aspergillus flavus and A. parasiticus were identified by macromorphological and molecular (calmodulin primer) features and screening of toxigenic and atoxigenic isolates was performed using of coconut-agar medium (CAM) and thin layer chromatography (TLC). This experiment was carried out in a complete randomized block design with four replications during the years 2012 to 2014.
 
Results
Out of 233 collected isolates of Aspergillus flavus clade, 221 and 12 isolates belonged to Aspergillus flavus and A. parasiticus, respectively. Frequency of toxigenic isolates of
A. flavus and A. parasiticus was 86.5, 87.5, 88.7% and 80, 75, 100% in cracked, intact fruits and soil, respectively. Based on the results of the combine analysis, the last irrigation treatments 30 and 25 days before harvest have the lowest and the last irrigation treatments 10 and 5 days before harvest have the highest population of Aspergillus flavus clade in the soil, intact and cracked fruits which showed significant difference with each other. Using of manure significantly increased the population of Aspergillus flavus clade, so that this increase in cracked, healthy fruits and soil in 2013 was 66, 83 and 114%, respectively.
 
Conclusions
The results of the present research showed that reduction of the interval between the last irrigation and the time of fruit harvesting (high moisture percentage of the surface soil of the orchard) and the using manure can increase the population of Aspergillus flavus clade in the soil, intact and cracked pistachio fruits. Increasing of the fungal population is accompanied by the frequency of toxigenic isolates of Aspergillus flavus and A. parasiticus. Adjusting the irrigation cycle, so that the last irrigation is 25 to 30 days before the fruit harvesting, can reduce the possibility of contamination of the pistachio kernels to Aspergillus and aflatoxin.

کلیدواژه‌ها [English]

  • Aflatoxin
  • Toxigenic isolates
  • Manure

مراجع

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مجله بیوتکنولوژی کشاورزی
دوره 14، شماره 4
دی 1401
صفحه 157-180

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