Research Paper I H J
Indian Horticulture Journal; 5(3/4): 86-92, July-December (2015) ©Indian Society of Advanced Horticulture ISSN: 2249-6823 DI: 179-15-IHJ-2208-2015-19
Application of a Steroidal Hormone Affects Fruit Yield and Quality in ‘Ataúlfo’ Mango (Mangifera indica L.) J F Aguirre-Medina, F Hernández-Hernández, A Sandoval-Esquívez*, L ArévaloGalarza**, A Ley De Coss and M R Gehrke-Velez Universidad Autónoma de Chiapas, Facultad de Ciencias Agrícolas, Huehuetán, Chiapas, México *Campo Experimental Rosario Izapa, INIFAP, México **Colegio de Posgraduados, Campus Montecillo, Texcoco, México e-mail: malcg.velez@gmail.com Received: 22 August 2015; Revised accepted: 04 November 2015
ABSTRACT In order to evaluate the effect of varying foliar applications of the brassinosteroid (BR) (CIDEF-4) prior to flowering on Mangifera indica L. cv ‘Ataulfo’ and to associate it with fruit quality and yield, the present study was conducted from September 2013 to May 2014 in a commercial orchard near Tapachula, Chiapas, Mexico. Treatments were: T1: untreated control with potassium nitrate (KNO2 2%) applied, T2: KNO2 2% plus Br (4g), T3: KNO2 (2%) plus 4 G of Br applied in two equal portions at different dates, (T4) KNO2 (2%) plus 4 G of Br applied in three equal portions at different dates. Each treatment was repeated ten times and one tree was considered an experimental unit. Flowering and fruiting variables were evaluated in the field and post harvest variables in the laboratory. Flowering was registered in ten inflorescences in each tree. Results indicate varying response to Br applications and to its effect on morphological and physiological yield variables. Yield per tree was increased with one Br application and flowering and fruit let presence increased with two applications. Postharvest fruit quality was enhanced showing more firmness and greater pectinmethylsterase activity and Brix readings. Fruits harvested from Br treated panicles showed greater weight loss. Key words: Brassinosteroids, Hormones, Ataulfo mango, Aminosteroids, Fruit yield Demand for „Ataúlfo‟ mango has increased in the national and international markets and this has favored its expansion, especially in the Pacific regions of Mexico and in other regions of central and South America. In Chiapas 27 691 hectares are reported, of which 62% are located in the Soconusco region (SIAP 2014) where it was first discovered. Decreases in yield are being observed due to diverse causes attributed to improper management and to environmental factors. Current results in plantations are reflected in high floral abortion rates, premature fruitlet drop and the presence of parthenocarpic fruits or “nubbins”. In recent years the application of diverse hormones such as brasinosteroids (Brs) (Chai et al. 2013) in low concentrations has been considered in Indian Horticulture Journal 5(3/4)
order to reduce the effects of stress in different plant organs (Nuñez and Mazorra 2001) and these have shown enhancement of growth and reproduction under adverse biotic and abiotic conditions (Krishna 2003, Bajguz and Hayat 2009). Among these are low and high temperatures (Nuñez et al. 2009). Regulatory activity seems to be due to the influence of these hormones on metabolic processes related to photosynthesis, nucleic acid and protein biosynthesis (Sasse 2003). Some Brs analogues which are similar to natural Brs have been shown to be useful in agriculture (Zullo et al. 2003). These compounds have a wide range of activity in protecting plant metabolism when these are under stress conditions and consequently increase their yield (Krishna 2003), through the 86
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Aguirre-Medina et al. 2015 and on November 14th. A total of four treatments were applied considering one tree as an experimental unit. Each tree was divided in four equal parts according to the cardinal points and each quadrant received the corresponding treatment clockwise. Treatments were applied in the morning from 7:00 to 9:00 a.m. with a 20 L. motor sprayer until trees were soaked to dripping point. Sprayed area was protected with plastic film to avoid spraying other parts of the tree. Morphological and physiological variables were recorded quarterly. Trees were harvested when the majority of fruits attained physiological ripeness. Once fruits were picked, these were placed in plastic crates and weighed with a digital platform weight (Ohaus DS20 L. USA) with a 100 kg capacity. Yield of each tree was registered separately. Postharvest variables during the fruit ripening process were evaluated for 10 days at two-day sampling intervals and weight loss was registered every day for nine days. Each fruit was considered an experimental unit. Response variables, Floral initiation. Once potassium nitrate was applied, flowering was recorded at each treatment. Panicle size: In ten trees, tenin florescences per tree were measured in cm. with a metric tape. Number of flowers was counted in each of ten inflorescences per tree in ten trees. The number of fruitlets in the “marble size” stage was recorded in each treatment in ten panicles in ten trees. Number of fruits was counted in each of ten panicles per tree in ten trees and the average number of fruits per panicle was calculated. Mango fruit weight was recorded using a granatory balance (Ohaus Adventurer Pro, USA) in ten panicles per tree in ten trees. Diameter was determined with a digital Vernier (AUTOTECTM China) in both fruit poles in ten fruits per tree in ten trees every fortnight for 67 days. Yield in t.ha-1. This variable was determined on the basis of weight of fruits produced per treatment per repetition. Firmness was determined by puncture measuring the force needed to penetrate the pulp with a texturometer (Chatillon, FDV-30 USA) with a 7 mm conical point. In each case one cm of the epicarp was eliminated on the sides opposite to the medium part. Thus two measurements were obtained per fruit. Values are reported in Newtons (N). Enzimatic activity of pectin methyl-esterase (PME). The Ranganna (1979) method was utilized which consists of applying citric pectin to 10 g of pulp as a substrate at pH 7.5 for 30 minutes. Number of methoxyl groups undoubled by the enzyme is quantified considering total soluble solids (oBrix) and sample weight. Total Soluble Solids (SST) (oBrix). Was determined with a digital refractometer (ATAGO Pallete Model PR-32α (0- 32%; USA) following the
increase of activity within the antioxidant route (Yin et al. 2008). The beneficial effects of Br‟s have been demonstrated in different crops such as vegetables, legumes, cereals, fruits and oilseeds (Janeczko et al. 2010, Hayat et al. 2012) in addition to enhancing crop quality (Peng et al. 2004). The object of this study was to evaluate the effect of the brasinosteroid CIDEF-4 using different foliar applications before and after flowering in „Ataúlfo‟ mango and to relate them to fruit yield and quality.
MATERIALS AND METHODS The field experiment was conducted at the “La Norteña” experimental site of the “Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP)” located at Km. 20.5 of the TapachulaPuerto Madero highway in the State of Chiapas. Location is 14°00´ - 14° 30´ N Lat and 92°00´-92°30´ W Long. Altitude is 40 m. above sea level. Laboratory research was conducted at the post crop physiology laboratory of the Postgraduate College Fruit program in Montecillo, State of Mexico, Mexico. Soil has a loamy clay texture pH 6.53 (1:2 H2O), O. M. 1.9%, NO-3 12.0 (mg.kg-1), NH4+ 14 (mg.kg-1), P 13 (mg.kg-1 Olsen), Fe2+ 21.9 (mg.kg-1), Mn2+ 19.8 (mg.kg-1), Zn2+ 9.1 (mg.kg-1), B 6.1 (mg.kg-1), K+ int. 289 (mg.kg-1), Ca2+ 2863 (mg.kg-1) y Mg2+ 185 (mg.kg-1), CE 0.07 (Dsm-1 a 25°C) and a 2% slope, superficial drainage is medium with slow infiltration and a 1.53 apparent density. Climate is warm sub-humid with rains in summer. Two dry seasons occur, one at midyear known as the “Canicula” (July-August) and the other from November to May. Climate according to Garcia (1987) is Aw (w”) i g with an annual rainfall of 1200 to 1500 mm and a mean annual temperature of 26ªC. The experiment was initiated with deweeding and sanitary practices (fruit removal), sanitary pruning of trees and fertilization once the previous year`s crop was removed in mid 2013. Nitrate was applied to all trees on October ninth of the same year. Steroidal hormone CIDEF-4 (Natura del Desierto S.A.) is a soluble nontoxic presentation compatible with fertilizer, fungicides and insecticides and a steroidal content of 80% (according to label information) was applied to the foliage on the indicated date. Treatments evaluated were: 1) Control (Potassium nitrate (KNO3) 2%); 2) Two g of Brasinosteroid /hectare applied on October 16; 3)Four g of Brasinosteroid /hectare applied in two equal parts on October 16th and 30th; 4) Six g of Brasinosteroid applied in three equal parts on October 16th and 30th Indian Horticulture Journal 5(3/4)
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Steroidal Hormone Affects Fruit Yield and Quality in ‘Ataúlfo’ Mango flower increase in „Ataúlfo‟ mango. In banana Jeyakumar et al. (2003) mention an increase of flowers upon application of (Br). Furthermore, Müssig (2005), noted that brassinosteroids influence ramification and flowering through modulation of the metabolism pattern and nutrient distribution. Largest number of marble-sized fruitlets was noted 33 and 11 days after the first and second application of (Br) respectively and similarly, the largest number of marble-sized fruitlets was counted 33 and 11 days after the first and second application of (Br) respectively. Under this condition, the largest number of these occurred when the hormone was applied in two applications. Increase was 45.9% higher in comparison with the control treatment and 52.2% when only one application of (Br) was made. There were no statistical differences due to the high coefficient of variation (Table 1). In cucumber, Fu et al. (2008) note the positive influence of (Br) during the initial stages of fruit development. Greatest number of fully developed fruits occurred when sprayed with 2 g of (Br). The remaining treatments with four and six g of (Br) showed 3.7 and 5.7 less mangoes respectively and represented 18.1 and 28.0% with regard to the 2 g. (Br) treatment.(Br) treatment in Passifloraedulis increased the number of fruits per plant in comparison with the untreated control (Gomes et al. 2006). In this respect, Susila et al. (2012) suggest that increased yield in fruit trees could be related to an increase in carbon assimilation and protein biosynthesis in trees sprayed with the hormone. Average fruit weights for ten fruits harvested at physiological maturity did not show significant statistical difference (P ≤ 0.05). However, greater fruit weight was obtained when sprayed three times with (Br). Wang et al. (2004) reported an increase in weight in oranges when brassinoid was applied. (Br) concentration also influences average fruit weight as observed in watermelon, probably due to an increase in cell division as compared to untreated plants (Susila et al. 2012).Fruits grew more in equatorial and polar diameter when trees were sprayed once with (Br) (Fig 1). Similar results have been found in other crops such as papayas (Mazorra et al. 2013). These authors report that (Br) effect caused an increase in fruit weight, diameter and size and increase in these values was directly related to spray rate. In our case in mangoes, the reverse was true. Largest mangoes were found when the lowest (Br) rate was applied. Fruit number per tree was increased and the largest amount occurred in the treatment where two g of (Br) were applied. Similar results have been found in other crops. Gomes et al. (2006) mention a 65% increase in yield in Passifloraedulis when treated with
AOAC methodology (1990). Five grams of fruit were squeezed in a mesh and placed in the refractometer. Results are expressed in oBrix. Titratable acidity (TA) was determined by the AOAC (1990) methodology which consists of liquefying 10g of pulp in 50ml of distilled water. A 5 ml aliquot of the solution was taken and titrated with NaOH (0.1N). Results are expressed as malic acid percentage. Weight loss was determined during nine days with a digital balance (Model EY-2200 A), and was considered as the individual initial and final weight of four fruits during the ripening process. Values are expressed in percent. Statistical analysis: Effects of treatments were determined by variance analysis for each variable using the PROC ANOVA procedure. Medium comparison was then carried out (Tukey P ≤ 0.05) using the Statistical Analysis System, version 8.1 (SAS 1999-2000) and plotted with the Sigma Plot ver. 10.0 for Windows.
RESULTS AND DISCUSSION Field values found related to floral initiation, panicle size, number of flowers per panicle and number of marble-sized fruitlets represent the results of Potassium nitrate application plus two applications of (Br). Data taken for these variables did not include final application of (Br) in the three-application treatment (T4) with the hormone. Initial potassium nitrate application was done on October ninth and the first and second hormone applications were carried out on October 16th and 30th. Counting of variables was done on November 13th, prior to the third application, realized one day later. In all treatments floral initiation occurred five or six days after potassium nitrate (KNO3) (2%) application and overall flowering was observed on November 13th. (Br) applications did not induce changes in this variable. Largest panicles occurred with the application of potassium nitrate alone and when the same treatment was sprayed with two applications of (Br). These results were statistically different (P ≤ 0.05), (Table 1). Previous growth can be enhanced by the stimulating effect of cellular division rates, such as occurred in isolated Petunia hybrid (Ho 2003) protoplasts in combination with cell division and leaf expansion (Zhiponova et al. 2013). Number of flowers per panicle did not show any outstanding effect with applications of (Br) (Table 1). One would not expect 12% decrease in flowers per panicle versus control when treated with two g of (Br). However, when foliage was sprayed twice, this variable increased 8% more than the untreated control. This effect suggests that two applications of (Br) favoured Indian Horticulture Journal 5(3/4)
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Aguirre-Medina et al. 2015 (Br) Biobras 16. Susila et al. (2012) reported yield increase in watermelon when sprayed with 0.1 ppm (Br) at the two- and four- leaf stage. Pipattanawang et
al. (1996) reported yield enhancement in strawberries and Peng et al. (2004) noted this in litchi (Nephelium litchi) fruits.
Table 1 Mean comparison of fruit size, number of flowers, “marble” fruit, fruits per panicle and average weight in „Ataúlfo‟ mangoes with different amounts of brasinosteroid in the coast of Chiapas, Mexico 2014 Panicle size Flowers Marble fruits Fully developed Fruit weight Fruit number Treatment (cm) panicle1 panicle-1 fruits panicle-1 (g fruit-1) per tree-1 Y KNO3 32.08±1.22 a 1000±153 a 18.5±2.17a 3.58±0.34 ab 314±14.0 a 812x KNO3 + 2g BrZ 26.95±0.99 b 879±132 a 17.7±6.46a 4.08±1.20 a 305±14.8 a 1060 (+30.5%) KNO3 + 4g Br 34.86±1.72 a 1084±144a 27.0±2.41a 3.34±1.09 ab 335±19.0 a 880 (+8.3%) KNO3 + 6g Br 2.94±0.19 c 361±19.7 a 960 (+18.2%) CV % 13.5 45.8 60.4 32.8 16.4 z
Br (Brasinoesteroid) Values with the same letter within each factor and column are equal according to tukey test P≤0.05. CV variation coefficient X Control related Y
Fig 1 Equatorially polar growth dynamics of ‘Ataúlfo’ mango in the coast of Chiapas, Mexico. Values are averages of 10 fruits per tree in ten trees during 67 days ± standard error
Fig 2 Weight loss of ‘Ataúlfo’mangoes with different amounts of Brassinosteroid in the coast of Chiapas. Values are averages of four fruits per tree. Verticalline indicates ± standard error
Mango firmness was affected significantly (P ≤ 0.05) at different moments during eight days of evaluation (Table 2). After day four, fruit firmness increased when (Br) application was increased from four to six g. At days six and eight greatest difference occurred when six g of (Br) were applied and at the final evaluation highest firmness value (not statistically different) was found when three sprayings of (Br) were applied. In other crops, such as papaya Mazorra et al. (2013) reported increased fruit firmness when (Br) was applied and Harindra et al. (2014) found greater firmness in grapes when sprayed with (Br). PME enzyme activity was more evident as of days eight and ten of the evaluation when (Br) was applied. In general, enzyme activity lessened in all treatments
as time advanced, although lowest value always occurred in the control as compared with treatments sprayed with (Br) (Table 2). (Br) application in lychee leaves prior to flowering increased PME activity (Peng et al. 2004). Total sugars found at the beginning of the evaluation increased as the amount of (Br) was increased. Highest value was found in fruits sprayed with six g of the hormone and was statistically different to the rest of the treatments (P ≤ 0.05). The evaluation made on the fourth day shows that highest concentration occurred in the treatment alone with the potassium nitrate application and was statistically different to the remaining treatments. Intermediate values were for treatments sprayed with two and six g of (Br). On the sixth day of sampling the same tendency recurred in fruit sugar content presented at
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Steroidal Hormone Affects Fruit Yield and Quality in ‘Ataúlfo’ Mango the start of the evaluation. On the eighth day, the highest value occurred in the 4 g of (Br) treatment. At the end of the evaluation, higher sugar content was found in the treatment alone with potassium nitrate applied. In general, results suggest more consistent response regarding average sugar content in treatments with potassium nitrate applied and when four and six g of (Br) were applied. Fruits treated with only potassium nitrate showed the highest initial amounts of TSS after two and six days storage as compared with treatments sprayed once and twice with (Br). As fruits ripened the amount found in the potassium nitrate treatment was 17.4ºBrix and for treatments where two g of (Br) were applied three times, values fluctuated
from 20.07 to 21.17 °Brix for samples taken after eight and ten days storage (Table 2). Increase in TSS content not influenced by (Br) concentration has been observed in watermelon (Susila et al. 2012). Gomes et al. (2006) have confirmed that there is a higher TSS content in Passiflora edulis in comparison with the untreated control when (Br) is applied. Mazorra et al. (2013) reported that in papaya Total soluble solids (TSS) in the fruit increased when (Br) was applied. Regarding acid content of the two following six days storage, highest values were found in treated trees where different (Br) applications were made. In almost all cases fruit acidity was higher when treated with (Br) (Table 2).
Table 2 Quality variables during „Ataúlfo‟ mango ripening sprayed at pre-flowering and pre-harvest with brassinosteroid and stored at room temperature (22 ± 2°C) 2014 Time Firmness PME activity Total sugars Total soluble solids Titrateable Treatments (days) (N fruit-1) (mg methoxyl g-1) fruit-1 (°Brix fruit-1) acidity fruit-1 z KNO3 15.28±0.45 ab 3.43±0.27 b 0.187±0.004 c 10.8±0.70 a 3.42±0.03 b Z KNO3 + 2 g Br 13.23±0.72 b 3.79±0.26 ab 0.202±0.002 c 9.6±0.22 a 4.32±0.04 b 2 KNO3 + 4 g Br 16.23±0.65 a 4.65±0.19 a 0.220±0.005 b 6.0±0.15 b 4.25±0.14 a KNO3 + 6 g Br 14.67±0.35ab 3.69±0.14 b 0.280±0.006 a 6.8±0.31 b 3.96±0.18 a CV % 9.3 11.6 11.6 6.06 6.06 KNO3 6.87±0.39 b 3.33±0.20 a 0.41±0.006 a 16.2±0.50 a 3.18±0.11 c KNO3+ 2 g BrZ 6.51±0.52 b 4.04±0.36 a 0.27±0.006 b 12.0±0.09 b 3.87±0.18 b 4 KNO3 + 4 g Br 11.67±0.78 a 3.66±0.16 a 0.20±0.004 c 8.3±0.15 c 4.70±0.06 a KNO3 + 6 g Br 12.67±0.39 a 4.07±0.29 a 0.27±0.008 b 9.1±0.11 c 4.52±0.19 a CV % 14.2 14.2 14.2 5.59 7.36 KNO3 1.24±0.07 ab 2.79±0.36 a 0.21±0.005 c 18.0±0.22 a 2.06±2.02 b KNO3+ 2 g BrZ 1.45±0.15 ab 3.42±0.17 a 0.30±0.009 b 17.8±0.21 a 2.56±0.02 a 6 KNO3 + 4 g Br 1.21±0.12 b 3.30±0.29 a 0.28±0.006 b 16.2±0.23 b 2.76±0.09 a KNO3 + 6 g Br 1.70±0.12 a 3.34±0.05 a 0.42±0.009 a 18.1±0.11 a 2.76±0.07 a CV % 21.6 15.7 15.7 2.31 5.14 KNO3 0.92±0.13 b 1.48±0.14 b 0.25±0.008 b 17.42±0.19 b 2.54±0.03 a Z KNO3+ 2 g Br 1.09±0.03 ab 2.02±0.36 b 0.19±0.002 c 16.97±0.30 b 2.55±0.05 a 8 KNO3 + 4 g Br 1.14±0.13 ab 3.24±0.23 a 0.33±0.002 a 18.32±0.25 b 2.56±0.14 a KNO3 + 6 g Br 1.45±0.16 a 3.19±0.28 a 0.18±0.00 c 21.17±0.49 a 2.52±0.04 a CV % 26.9 21.8 21.8 3.60 6.71 KNO3 0.59±0.01 a 0.36±0.02 c 0.35±0.022 a 18.05±0.47 b 1.17±0.03 b Z KNO3+ 2 g Br 0.61±0.02 a 0.41±0.01 c 0.25±0.016 b 19.80±0.24 a 1.27±0.03 b 10 KNO3 + 4 g Br 0.57±0.04 a 1.12±0.07 a 0.19±0.004 b 17.92±0.30 b 1.97±0.04 a KNO3 + 6 g Br 0.65±0.03 a 0.59±0.02 b 0.25±0.014 b 20.07±0.24 a 1.38±0.10 b CV % 12.7 13.1 13.1 3.50 8.82 Weight loss was slightly higher in treatments sprayed with (Br) (Fig 2). Untreated control lost an average of 1.5% daily with two g of (Br), 1.6% with four g of (Br) and 1.7% daily weight loss when applied Indian Horticulture Journal 5(3/4)
with six g of (Br). Weight loss began to increase at day three for treatments sprayed with four and six g of (Br). Mango fruit weight loss is due principally to transpiration. Untreated control lost 12.4% when 90
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Aguirre-Medina et al. 2015 applied with 2 g of (Br), 13.6% with 4 g of (Br) and 13.9% with 6 g. Exogenous applications of (Br) in vegetative tissue induce ethylene production (DeGrauwe et al. 2005, Arteca and Arteca 2008). In some fruits, such as mangoes and strawberries, (Br) is present in small quantities and may not be critical to ripening (Zaharah et al. 2011, Symons et al. 2012), but exogenous applications may induce ethylene production (Zaharah et al. 2011). In the Kensington Pride variety, Zaharah et al. (2011) found that ethylene production and peak respiration occurred at day four of ripeness and at this time (Br) in the fruits occurred
in trace quantities. However, exogenous application of Epi-BL (45 y 60 ng g-1) promoted fruit ripeness. From the investigation it was observed that varying response to brasinosteroid application and its effect on morphological and physiological yield variables was found. Fruit yield per tree increased with one application of the brasinosteroid and two applications enhanced flowering and presence of small fruits. Fruit quality was enhanced as regards firmness during the first days of evaluation as were pectin methylsterase activity and °Brix. (Br) sprays applied to flowers induced greater weight loss.
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