تا ثير بعض المعاملات الحيوية والكيماوية بعد الحصاد على تخزين ثمار البرتقال

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1 استمارة معلومات الرساي ل التى تمت مناقشتها الكلية : الزراعة القسم: بساتين الفاكهة 1 2 الدرجة العلمية : دكتوراه بيانات الرسالة : عنوان الرسالة باللغة العربية : 3 تا ثير بعض المعاملات الحيوية والكيماوية بعد الحصاد على تخزين ثمار البرتقال عنوان الرسالة باللغة الا جنبية : EFFECT OF SOME POST HARVEST BIOLOGICAL AND CHEMICAL TREATMENTS ON STORAGE OF ORANGE FRUITS التخصص الدقيق : بساتين - الفاكهة تاريخ المناقشة : بيانات الطالب : الاسم : زينب ا حمد زكى ا حمد الجنسية : مصرية النوع : ا نثى العنوان :المعادى-شارع ا حمد زكى 30 تليفون جهة العمل : معهد بحوث البساتين 4 المشرفون على الرسالة : القسم الاسم الفاكهة ا.د.محمد رضا بركات د. محمد ا حمد عيسى ا.د. محمود على ا حمد الفاكهة تداول الفاكهة شارع المشير ا حمد ا سماعيل رقم الفاكس : البريد الا لكترونى : Zeinab_mona@yahoo.com الكلیة الزراعة الزراعة معهد بحوث البساتين الجامعة القاهرة القاهرة مركز البحوث الزراعية 6/1

2 5 5 مستخلص الرسالة ) Abstract ( 1 باللغة العربية : بشرط ا لا يزيد عن 7 ا سطر ا جريت هذه الدراسة خلال موسمي , على ثمار البرتقال ا بو سره والفالنشيا بمعمل قسم بحوث تداول الفاكهه بمعهد بحوث البساتين مركز البحوث الزراعية فى محاوله لايجاد وسيله امنه وفعاله لمكافحة اعفان ثمار البرتقال اثناء الشحن او التخزين كبديل لاستخدام المبيدات الكيماويه. تم معاملة الثمار بالنقع في محلول فطر Trichoderma album وا خرى في محلول بكتريا megaterium, 2000 جزء فى المليون بتركيز 1500, 1000, 500, Bacillus تركيز 1500, 1000, جزء فى المليون لمدة 3 او 5 دقاي ق.كذلك تم معاملة مجموعة من الثمار بالايمازيليل كمعاملة كيماوية وكذلك تم تشميع فقط لمجموعة ا خرى من الثمار ) الكنترول). ثم تم تخزين الثمار المعاملة والغير معاملة على درجة (C 10 ورطوبة نسبية %). تم فحص الثمار كل ثلاثة ا سابيع لمتابعة التغيرات الطبيعية والكيماي ية في الثمار خلال فترة التخزين. وقد ا ظهرت النتاي ج ا ن المعاملة بكل من الفطر والبكتريا ا فضل من المعاملة بالمادة الكيماوية (الايمازيليل) والكنترول لمكافحة الاعفان التي تصيب الثمار خلال فترة التخزين ويرجع ذلك ا لى ا ن هذه المواد الحيوية المستخدمة مضادة طبيعيا للكاي نات الممرضة بوضعها على سطوح الثمار لتضاد الكاي نات المسببة لامراض ما بعد الحصاد ومن ثم يمكن استخدام اى من المركبات المشار اليها سابقا غمرا كبديل للايمازيليل مع العلم ا ن هذه المواد امنه صحيا ويمكن استخدامها كبديل للمواد الكيماوية ) الايمازيليل) والتى ثبت تركها اثرا متبقيا داخل الثمرة مما يضر بصحة المستهلك. ويمكن التوصية بنقع ثمار البرتقال فى محلول Bacillus megaterium بتركيز 1500 جزء فى المليون ولمدة خمسة دقاي ق حيث تفوق على المعاملات الاخرى فى المحافظة على الثمار بجودة عالية ولاطول فترة ممكنة. Trichoderma Bacillus الكلمات الداله: (ثمار الموالح تخزين بعد الحصاد ايمازيليل) 6/2

3 5 2 باللغة الا جنبية : بشرط ا لا يزيد عن 7 ا سطر Key Words : (Citrus fruits,storage, postharvest, Trichoderma, Bacillus,Imazalil.) This study was carried out during two successive seasons & on Washington navel orange and Valencia orange at Fruit Handling Department, Horticulture Research Institute, Agriculture Research Center, Giza, Egypt. Fruits were subjected to the following treatments: dipped in solutions of 500,1000,1500,2000 ppm Trichoderma album or Bacillus megaterium for three or five minutes for each treatment or in 1500 ppm Imazalil, (the traditional packaging house treatment), or without any other treatment (waxing only) as control. Fruits were stored at 10 C and % RH. The results showed that, dipping postharvest treatments with either Trichoderma album or Bacillus megaterium had superior effect on reducing orange fruit rotting and keeping quality during storage in comparison with either untreated or treated with Imazalil. However, dipping postharvest treatments with Bacillus megaterium solution at 1500 ppm for 5 minutes was the best treatment in order to keep orange fruit postharvest quality. 6/3

4 ا هم النتاي ج التطبيقية التى تم التوصل ا ليها : 6 ) لا تزيد عن سطرين لكل منها ( 1 يمكن ا ستبدال المركب الكيماوى الا يمازيليل المستخدم فى محطات تعبي ة ثمار الموالح 6 والذى ثبت تركه ا ثرا متبقيا داخل ثمار البرتقال مما يشكل خطورة واضحة على صحة المستهلك بالمستحضر البكتيرى Bacillus megaterium 2 كانت ا فضل النتاي ج المتحصل عليها هى نقع ثمار البرتقال فى المستحضر البكتيرى 6 بتركيز 1500 جزء فى المليون لمدة خمس دقاي ق وذلك Bacillus megaterium للمحافظة على ثمار البرتقال من الاصابة بالاعفان خاصة العفن الاخضر والعفن الازرق. 6/4

5 ما هى الجهات التى يمكن ا ن تستفيد من هذا البحث : 7 ) اذكر هذه الجهات مع شرح ا همية البحث لهذه الجهة بما لا يزيد عن ا ربعة سطور لكل جهة) ) -2 7 محطات تعبي ة ثمار الموالح) حيث يتم فى هذه المحطات فرز ثم غسيل الثمار ثم بعد ذلك يتم معاملتها بالمطهرات الفطرية مثل الايمازيليل والثيوبندازول والتى ثبت تركها اثر متبقى ليس فقط على قشرة الثمرة ولكن هذا الاثر يتغلل الى قلب الثمرة مما يعرض الشحنة للرفض تجاه الدول المستوردة للبرتقال ومن جهة ا خرى تشكل النسبة المتبقية من هذه المطهرات داخل الثمرة خطورة واضحة على صحة المستهلك لذلك توصى هذه الدراسة باستخدام المستخلص البكتيرى الباسيلس ميجاتيريم بتركيز 1500 جزء فى المليون غمرا ولمدة خمس دقاي ق بدلا من الايمازيليل حيث انه امن من الناحية الصحية واقتصاديا من الناحية التجارية. 8 هل توجد علاقة قاي مة با حدى هذا الجهات : فى حالة نعم اذكر هذه الجهات : لا ما هى طبيعة العلاقة : مشروع بحثى تعاون ا كاديمى ( : مشروع ممول من جهة ثالثة ) اذكر ما هى ( ) ا خرى تذكر 6/5

6 ب( ج( هل توافق على التعاون مع جهات مستفيدة من خلال الجامعة : 9 نعم (ا ) ( ( لتطبيق البحث : وكذلك محاولة ا ختبار المركبات الحيوية محل الدراسة على محاصيل ا خرى. لاستكمال البحث : ا خرى ( ) تذكر 10 هل تم نشر بحوث مستخرجة من الرسالة فى مجلات ا و مو تمرات علمية ) تذكر مع جهة النشر و المكان و التاريخ ( 10 1 نعم تم نشر بحث فى مجلة Journal of Horticultural Science & Ornamental Plants 4 (1): 50-57, 2012 IDOSI Publications, 2012 ISSN بعنوان Effect of Using Some Biological Post Harvest Treatments on Storability of Washington Navel Orange Fruits Compared with Imazalil Post Harvest Chemical Treatments تا ثیر إستخدام بعض المعاملات الحیویة بعد الحصاد على القدرة التخزینیة لثمار البرتقال أبو سرة مقارنة بالایمازیلیل كمعاملة كیماویة. لا 11 هل سبق التقدم لتسجيل براءات اختراع ) لا تذكر مع الجهة و المكان و التاريخ ( 12 هل توافق على ا عطاء البيانات المذكورة فى هذه الاستمارة لجهات ا خرى نعم توقيع المشرفين : توقيع الطالب : زينب ا حمد زكى ا حمد ا.د. محمد رضا بركات ا.د. محمد ا حمد عيسى ا.د. محمود على ا حمد وكيل الكلية ) المعهد ( للدراسات العليا و البحوث التاريخ : 6/6

7 ACKNOWLEDGEMENT Firstly my unlimited thanks to "Allah" Lord of the world. I wish to express my thanks and pay my respect to Dr. Mohamed Reda Barakat, Professor of Pomology, Cairo University for his supervision, criticism and all his efforts and huge contribution to the success of this work. All my regards, thanks and respect to Dr. Mohamed Ahmed Eissa assistant Professor of Pomology, Cairo University for his supervision guidance, his efforts and sincere assistance during this work. Deep gratitude is due to Dr. Mahmoud Ali Ahmed Head Research, in Fruit handling of Horticulture Research Institute in Agriculture Research Center, Giza, Egypt. Grateful appreciation is also extended to all staff members of Pomology Department, Fac. Agric., Cairo University. Special deep appreciation is given to my family and friends. Also, I feel deeply grateful to all staff members of Horticulture Research Institute, Agriculture Research Center, Giza, Egypt who helped me without mention any name to avoid forgetting anyone.

8 DEDICATION I dedicate this work to whom my heart felt thanks; to my late mother. Also I dedicate this work to my father and my brothers for their patience and helps, as well as to my friends and my colleges.

9 INTRODUCTION Orange industry is important for Egyptian National Income. Orange Planted area in Egypt reached Feddans in The producing area reached feddans producing tons of orange fruits according to the statistics of Ministry of Agriculture, Egypt (2010). Although orange occupies the greatest planted area among all citrus grown fruit area in Egypt, the exportation of fresh orange fruits to foreign markets is still limited compared with the produced quantity. Therefore, any effort directed towards maintaining fruit quality and reducing postharvest losses is important for increasing our National income. Citrus is one of the major commercial fruit crops that are widely consumed as both fresh fruit and juice. Its global demand is attributed to its high vitamin C content and its antioxidant potential (Gorinstein et al. 2001). Unfortunately, citrus is attacked by several plant pathogens that affect its fruit quality. In developing countries, where protection and proper handling of fresh fruit is inadequate, losses during transit and storage can represent over 50 % of the harvested crop (Eckert and Ogawa 1988 Wisniewski and Wilson 1992). Major postharvest losses were associated with a range of pathogens. They include green and blue mould caused by Penicillium spp., gray mould caused by Botrytis cinerea Pers ex Fr (Agrios 1997). However, it is often reported that the importance and impact of these pathogens on the citrus industry differ from country to country. Therefore, it is important for a country to first 1

10 determine the spectrum and relevant importance of the pathogens involved in postharvest decay. In order to reduce postharvest losses, synthetic fungicides are applied either pre- or postharvest. However, the application of synthetic chemical compounds to control postharvest diseases often result in chemical residues on food that may affect human health (Norman 1988) and development of resistant pathogens (Wilson and Wisniewski, 1989). Eckert (1988) explained about the residual effects of Imazalil that,six Imazalil tolerant isolates tested were cross resistant to propiconazole, fenarimol, flusilazole, penconazole, myclobutanil,prochloraz, diclobutrazole and bitertanol, all of which inhibit the C-14 demethylation reaction in sterol biosynthesis. Tests showed that the standard Imazalil spray treatments ( ppm) which give residues of 1-2 ppm in citrus fruit will not control either infection or sporulation by tolerant penecillium digitatum biotypes. Tomlin (1997) added that, Thiabendazole and Imazalil are both mainly used as postharvest pesticides; this means that they should be located on the peel of oranges, which was also confirmed by our results. At the same time a small amount of Imazalil was found in the pulp as well. This fact can be explained by diffusion of Imazalil through the peel of oranges. The tendency of diffusion can be explained by high polarity of Imazalil. The inside of orange is more polar because of the high content of water. This explains the tendency of polar Imazalil to diffuse through the orange peel. On the other hand many authors demonstrated that, Imazalil is not only found on the peel 2

11 of Apple, but also in the pulp. This fact supports the conclusion that Imazalil has a high diffusion rate. These findings show that even if the pesticides are used in the postharvest, they may diffuse through the peel of fruits and are also found in the pulp. Bolognesi and Morasso (2000) reported that, pesticides are widely used in agriculture to fight weeds, moulds, and pests for increasing productivity. Besides this positive effect pesticides pose health-risk to consumers. Guidance documents on residues analytical methods, reported that, the concentrations of pesticides residues in many products, including fruit and vegetables, must be monitored and regulations such as SANCO/825/00. More over Anneil et al. (2007) studied pesticide residues in commercially available oranges and on evaluation of potential washing methods. The results showed that, pesticides which are widely used in agriculture to increase production or protect crop, are known to be potentially dangerous to consumers. Therefore oranges from local supermarket were analyzed by the LC-ESI-MS method to determine the presence of pesticides. Two pesticides - Thiabendazole and Imazalil were found in the oranges. Both are postharvest pesticides. Analyses showed that, these pesticides are located in and on the peel of oranges. Five different washing methods washing with cold water, with hot water, with soap, with dishwashing soap, and treating with ultrasound were evaluated for their efficiency in removing pesticide residues. They added that, none of the used methods were able to remove Imazalil totally. Washing with cold and hot water decreased the concentration by about one third, but using soap was ineffective. 3

12 From all the used methods, washing in ultrasonic bath was the most effective in removing Imazalil. Therefore, the development and use of alternative postharvest control options involving biological agents or natural plant extracts have become important since it is perceived as being environmentally safer and more acceptable to the general public (Janisiewicz and Korsten, 2002). Therefore, the objectives of this study were: 1. Reduce the percentage of fruit rot. 2. Replace the used chemical treatments by the biological treatments. 3. Identifing the best treatment and the best concentration. 4. Store the fruits for long term in good quality. 4

13 Name of Candidate: Zeinab Ahmed Zaki Ahmed Degree: Ph.D. Title of Thesis: Effect of Some Postharvest Biological and Chemical Treatments on Storage of Orange Fruits Supervisors: Dr. Mohamed Reda Barakat Dr. Mohamed Ahmed Eissa Dr. Mahmoud Ali Ahmed Department: Pomology Branch:- Approval: 30 / 6 /2012 ABSTRACT This study was carried out during two successive seasons & on Washington navel orange and Valencia orange at Fruit Handling Department, Horticulture Research Institute, Agriculture Research Center, Giza, Egypt. Fruits were subjected to the following treatments: dipped in solutions of 500,1000,1500,2000 ppm Trichoderma album or Bacillus megaterium for three or five minutes for each treatment or in 1500 ppm Imazalil, (the traditional packaging house treatment), or without any other treatment (waxing only) as control. Fruits were stored at 10 C and % RH. Fruits were statistical analyzed in two stages: the first stage concluded comparison of the different concentrations from the biological treatments by the control fruits. The second stage concluded comparison of the standard concentrations from the biological treatments by the chemical treatments (Imazalil). The results show in the first stage that, the fruits quality and storability increased with the increasing concentration and duration of the biological treatment compared with the untreated fruits (control) for 24 and 21 weeks during the first and the second season respectively. Fruits were analyzed for physical and chemical changes at 3 weeks intervals during storage periods.the results of the second stage show that, fruits treated with either Trichoderma album or Bacillus megaterium showed less decay and weight loss incidence during storage compared with the other fruits which were either treated with Imazalil or untreated at all (waxing only). Moreover, all the examined postharvest treatments significantly reduced physical and chemical properties deterioration rates of fruits during storage. Biological control postharvest treatments significantly reduced the decreasing of fruit juice%, color transition firmness, total acidity and ascorbic acid juice contents. On the other hand, these treatments increased total sugars,tss, TSS/Acid ratio contents of fruits.in brief, it s concluded that, dipping postharvest treatments with either Trichoderma album or Bacillus megaterium had superior effect on reducing orange fruit rotting and keeping quality during storage in comparison with either untreated or treated with Imazalil. However, dipping postharvest treatments with Bacillus megaterium solution at 1500 ppm for 5 minutes was the best treatment in order to keep orange fruit postharvest quality. Key words: Citrus fruits,storage, postharvest, Trichoderma, Bacillus,Imazalil

14 MATERIALS AND METHODS This investigation was carried out during two successive seasons ( ), ( ) at the Agriculture Research Centre - Hort. Research Institute - Fruit Handling Department, Giza Governorate, Egypt, on Washington Navel, Valencia oranges. The fruits were picked in 27 th Feb., 2008 and in 22 nd Feb., 2009 for Valencia oranges and 13 th Dec., 2008 and 10 th Dec., 2009 seasons for Navel oranges from a private farm, in El-Fayoum when they reached the maturity stage as determined by T.S.S./acid ratio ( ) and Juice percentage reached % (Kader, 1985). Fruits from each cultivar were packed in plastic boxes and immediately transported to the Fruit Handling Laboratory Hort. Res. Institute. In the laboratory, the fruits were divided into four groupes and treated as follows: 1. Control fruits were washing, draying and waxed. 2. Fruits were dipped in Imazilil (1500 ppm) for (3) minute, then dried and waxed. 3. Fruits were dipped in solutions of Trichoderma album for (3, 5) minutes with the concentrations 500, 1000, 1500, 2000 ppm,then dried and waxed. 4. Fruits were dipped in solutions of Bacillus megaterium for (3, 5) minutes with the concentrations 500, 1000, 1500, 2000 ppm,then dried and waxed. After that fruits of each group were stored in carton boxes at 10 C, % RH to determine the physical and chemical changes during storage. And then fruits were statistical analysis on two stages, 30

15 the first stage concluded the comparison between different concentrations from the biological treatments with untreated fruits (control), the others concluded comparison between the standard concentrations from the biological treatments with Imazalil (chemical treatments). Methods of determining fruit properties a. physical properties 1. Weight loss percentage It was determend by six fruits at each examination period were weighted and the difference between its weight at the beginning of the experiment and at the examination day were represented as weight loss percentage using the following equation, Weight loss % = weight loss (g) / Initial weight of fruits Decay percentage It was detwrmined by the weight of unmarketable fruits due to pathological disorders was determined and this percentage was calculated as follows, Decay % = Decayed fruit / Initial fruit weight Fruits firmness These values were determined using Lefra Texture Analyses at 5 mm depth and 0.2 mm /second, the pressure was on two sid of fruit, for measuring firmness of orange, this value was determined by taking the firmness value of six fruits from two sides for each cultivar and the average of the fruit was measured as (g/cm 2 ). 4. Peel color 31

16 These values were estimated in six fruits by using hunter colorimeter model DP 9000 (hue angel) were determined according to Mc Gjuire (1992). 5. Juice percentage It was determined by six fruits for each treatment represent three replicates were used. This percentage was determined by weighting the fruits and weighting the extracted using this equation Juice % = Juice weight / Fruits weight 100 b. Chemical properties: 1. Total soluble solids (TSS) percentage It was determined by abbe-digital refractometer. 2. Titratable acidity percentage It was determined as citric acid by titrating with a standard solution of sodium hydroxide (0.1 N), using phenolphthalein as an indicator, Acidity was calculated as mg/100 ml of the juice according to A.O.A.C. (1995). Acidity was calculated by this equation Acidity (g) citricacid / (100ml) juice = ml NaOH x 0.1 x / ml juice x Total soluble solids: titratable acidity ratio These values were obtained from dividing the percentage of total soluble solids by that of titratable acidity. 4. Ascorbic acid content (Vitamin C) It was determined by using 2,6-di chloro phenol in % phenol dye and 2% oxalic acid as a substrate. Ascorbic acid was calculated as mg / 100 ml of the juice (Lucass, 1944). 5. Total sugars contents It was determined by A.O.A.C,

17 c. Statistical analysis The obtained data in the two seasons of the experiment were statistically analyzed as factorial randomized complete design. Means were compared by the least significant difference test (L.S.D) at the 5% level of probability according to Silva, F. de A. S. e., Azevedo, C. A. V. de. (2009). 33