اثر تلقيح بذري قارچ Piriformospora indica و محلول پاشي پاكلوبوترازول بر مقاومت به تنش سرمازدگي در گياه لوبيا سبز (.Phaseoluse vulgaris L)

The effect of Piriformospora indica seed bio-priming and paclobutrazol foliar spraying on tolerance to chilling stress in green beans (Phaseolus vulgaris L.)

كاهش عملكرد ناشي از تنش سرمازدگي در گياهان گرمسيري و نيمه­ گرمسيري اهميت بررسي اين تنش و تيمارهايي كه مي­ توانند اثرات آن را كاهش دهند را نمايان مي­سازد. به­ منظور بررسي اثر قارچ اندوفيت Piriformospora indica و پاكلوبوترازول در القاي تحمل به تنش سرمازدگي در گياه لوبيا سبز (Phaseolus vulgaris L.)، آزمايشي گلخانه ­اي به ­صورت فاكتوريل بر پايه‌ي طرح كاملاً تصادفي در سه تكرار در گلخانه تحقيقاتي پژوهشكده­ ژنتيك و زيست‌فناوري كشاورزي طبرستان واقع در دانشگاه علوم كشاورزي و منابع طبيعي ساري اجرا شد. فاكتورهاي آزمايشي شامل تنش سرمازدگي در سه سطح (شاهد، سه و شش روز تنش سرمازدگي با دماي پنج درجه سانتي­گراد)، تلقيح قارچي در دو سطح (بدون تلقيح و تلقيح قارچ P. indica) و محلول­ پاشي پاكلوبوترازول در سه سطح (صفر، 40 و 80 ميلي­گرم در ليتر) بود. تلقيح قارچ در مرحله­ كاشت، محلول­ پاشي پاكلوبوترازول پنج و شش هفته و تنش سرمازدگي هشت هفته پس از كاشت اعمال گرديد. نتايج نشان داد كه تنش سرمازدگي به­ ويژه به مدت شش روزسبب كاهش وزن خشك اندام­هاي هوايي و ميزان كلروفيل a، b و كاروتنوئيد (به ترتيب حدود 19، 18 و 8 درصد) نسبت به تيمار شاهد گرديد. در تمام سطوح تنش، تلقيح قارچ P. indicaوزن خشك بوته را افزايش داد به‌طوري‌كه در تنش شديد، بيشترين تأثير (افزايش 65 درصدي) را داشت. محلول­ پاشي پاكلوبوترازول با غلظت 40 ميلي­گرم در ليتر، كلروفيل a، كاروتنوئيد و محتواي كلروفيل برگ را به ترتيب 5، 9 و 7 درصد افزايش داد. همچنين محلول­ پاشي پاكلوبوترازول و تلقيح قارچ سبب كاهش نشت الكتروليت شد. درمجموع قارچ P. indica و پاكلوبوترازول در تعديل اثرات تنش سرما و بهبود رشد گياه لوبيا سبز اثر مؤثري داشتند.

Introduction Green bean (Phaseolus vulgaris L.) is one of the four species of the genus Phaseolus with the highest protein among a variety of commercial plants (Nasri and Khalatbari, 2011(. This plant can be grown in different times in Mazandaran weather conditions (Motaghian et al., 2010). Green beans as well as other crops during their growth stages faced with environmental stresses such as chilling. Chilling stress referred to conditions as temperature between 0 and 10 ᵒC which resulted in damages to the plant (Baninasab, 2009). Chemical and biological inducers can be used to increase chilling stress tolerance of plants. Paclobutrazol (PBZ) is a member of the Tryazol families with growth regulating effect (inhibitors) and increased stress tolerance through changes in plant growth and morphological, anatomical and biochemical changes (Berova et al., 2002; Chaturvedi et al., 2009). Meanwhile, Piriformospora indica fungus is one of the soil microorganisms which form a symbiotic relationship with plants and could increase plant growth and resistance to environmental stresses throughout systemic changes or defense induction (Deshmukh et al., 2006). Hence, the present research was aimed to determine PBZ and P. indica efficiency in green bean tolerance against chilling stress. Materials and Methods The experiment was conducted under controlled conditions at Genetics and Agricultural Biotechnology Research Institute, Sari Agricultural Sciences and Natural Resources University in factorial arrangement based on completely randomized design with three replications in 2013. Treatments were three levels of chilling stress (control, three and six days exposure to five centigrade degree temperature), two levels of fungi inoculations (control and inoculation with P. indica) and three levels of PBZ (0, 40 and 80 mg/L). To inoculate the plants with P. indica, fungal spore suspension was prepared (5×105 spore/ml). The sprouted seed were soaked for four hours in fungal suspension and were sown in pots containing two kg of sterilized soil. PBZ was sprayed twice at two stages of five and six week after sowing. Then pots were transferred to growth chamber after eight weeks from sowing and then chilling stress was imposed. After 12 hours of stress, aerial dry weight, SPAD value, chlorophyll a, b and a+b, carotenoid content and electrolyte leakage were measured. The data were analyzed using SAS statistical software and significant differences between means were estimated using least significant difference (LSD) test at P<0.05 of probability level. Results and discussion Results showed that chilling stress particularly for six days decreased aerial dry weight, chlorophyll a and b and carotenoid content (nearly 19, 18 and 8%, respectively) as compared to the uninoculated control. In all levels of chilling stress, however, P. indica inoculation increased plant dry weight in which the most beneficial effect (up to 65%) was recorded in severe stress. Mean comparison of three factors indicated that at all levels of chilling stress and PBZ, P. indica increased leaf and whole plant dry weight at the range of eight to 280% and 0 to 143%, respectively. The maximum increase for leaf dry weight (2.68 g/plant) was recorded where inoculated plants grown in normal conditions in no applied PBZ. Furthermore, the highest value of aerial dry weight (3.42 g/plant) recorded in plants which inoculated both with P. indica and sprayed with 80 mg/L of PBZ under normal conditions. PBZ spraying at the rate of 40 mg/L increased chlorophyll a, carotenoid and SPAD value by 5, 9 and 7%, respectively. In inoculated plants, however, PBZ spraying decreased plant dry weight particularly at the rate of 40 mg/L (nearly 17%). PBZ spraying, also, reduced electrolyte leakage in which the maximum reduction (up to 20% lower than non-application) was obtained with 80 mg/L of PBZ. Conclusion In conclusion, the results indicated that P. indica inoculation along with Paclobutrazol spraying had a positive effect on chilling tolerance in green beans. Therefore, these factors can be used to induce chilling stress tolerance. However, further tests should also be applied to other growth stages of plant.